LOCUS       sequence-471814-        4917 bp    DNA     circular SYN 16-JUN-2025
DEFINITION  Express MycMax response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4917)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL [NlucP/MycMax-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..152
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      179..188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             185..697
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             701..820
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    872..993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(883..902)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     937..956
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1187..1544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1187..1378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1210..1230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1395..1530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1457..1476
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1575..2612
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2591..2599
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2636..2684
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2840..2857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3011..3599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3091..3110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3799..4590,4591..4659))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4764..4812
                     /note="synthetic polyadenylation signal"
     misc_feature    4826..4917
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4866..4885
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcagac cacgtggtca gaccacgtgg tcagaccacg
       61 tggtcagacc acgtggtcag accacgtggt cagaccacgt ggtcctcgaa gcttagacac
      121 tagagggtat ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaagc
      181 caccatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa
      241 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc
      301 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca
      361 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt
      421 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact
      481 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat
      541 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat
      601 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg
      661 agtgaccggc tggcggctgt gcgaacgcat tctggcgaat tctcacggct ttccgcctga
      721 ggttgaagag caagccgccg gtacattgcc tatgtcctgc gcacaagaaa gcggtatgga
      781 ccggcaccca gccgcttgtg cttcagctcg catcaacgtc taaggccgcg actctagagt
      841 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
      901 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
      961 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1021 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1081 ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct tccgctgatc tgcgcagcac
     1141 catggcctga aataacctct gaaagaggaa cttggttagc taccttctga ggcggaaaga
     1201 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1261 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1321 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1381 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1441 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     1501 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc gattcttctg
     1561 acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa aaatttctca
     1621 tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa gagagccgag
     1681 ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg
     1741 gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta cccatccccg
     1801 aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt agacgcgccc
     1861 aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg
     1921 ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc ttcggcccat
     1981 tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc gccattgctg
     2041 atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc agcgtagctc
     2101 aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc
     2161 atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg
     2221 actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc ttcttctggc
     2281 ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg catcccgagc
     2341 tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat cagctctacc
     2401 agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca
     2461 tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg agcgcagccg
     2521 tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg cccagtacac
     2581 gaccgcgcgc taaggaggta ggtcgagttt aaactctaga accggtcatg gccgcaataa
     2641 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgttcgaa ctagatgctg
     2701 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2761 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2821 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat
     3781 gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg
     3841 cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga
     3901 gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc
     3961 cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac
     4021 tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc
     4081 agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc
     4141 gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc
     4201 catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt
     4261 ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc
     4321 atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg
     4381 tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag
     4441 cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat
     4501 cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc
     4561 atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa
     4621 gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta
     4681 ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt
     4741 acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     4801 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
     4861 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctct
//
